Re: Get a plasmide - buy? share?

What he performed was a general DNA extraction, and a very crude one
at that. The majority of the DNA isolated in that prep was probably
nucleoid and not plasmid. I am not completely sure, but I would
estimate that the plasmid content would be very low. One thing that I
would worry about in his procedure would be nuclease activity. Using
something like TE buffer instead of water should help to deactivate
them before they chew up all the DNA. The chromosomal DNA here is a
large part of what is ultimately extracted. The dish detergent mostly
serves to disrupt the lipid membranes/lyse the cells.

For a plain old transformation, the plasmids don't have to be
extremely pure, but I imagine efficiency would drop off at a certain
point. However if you plan to do any analysis (electrophoresis,
sequencing, etc) or engineering on the plasmid, then you would need
the most pure form available. In this

On Dec 30, 4:43 am, Mega <masterstorm...@gmail.com> wrote:
> Wow.
> Have a look at that:http://www.youtube.com/watch?v=TZeL7WIkQC0
> He just uses a dish detergent and sodiumhydroxide (both of which I
> already have at home) and isopropanol  to conduct an alkaline -home-
> miniprep!!
>
> I immagine that the purity of the plasmids is not very high, maybe
> some 80-85% ??
>
> But anyway, is it neccessary for a transformaiton to have relatively
> pure plasmids? Some proteins and other substances (e.g. luciferin,
> sugar, etc.)  that are in the solution don't affect the would-be
> transformed bacteria, thay may consume them or not. No big deal?
>
> But, what happens to the chromosomal DNA in this process? Is it
> denatureted by the dish detergent??

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